Cell support composite and method for producing cell support composite

Patent metadata

Field	Value
Publication number	US-10870824-B2
Application number	US-201815970092-A
Country	US
Kind code	B2
Filing date	May 3, 2018
Priority date	Nov 10, 2015
Publication date	Dec 22, 2020
Grant date	Dec 22, 2020

Abstract

A cell support composite includes: a substrate formed of an artificial material; a laminin molecule or a fragment thereof which adheres to at least a part of the substrate; and a renal tubule epithelial cell or a renal tubule epithelial-like cell which is a culture cell attached to the substrate via the laminin molecule or the fragment thereof.

First claim

What is claimed is:

1. A cell support composite comprising:
a substrate formed of an artificial material;
a laminin molecule or a fragment thereof which adheres to at least a part of the substrate; and
a monolayer of a renal tubule epithelial cell or a renal tubule epithelial-like cell which is a cultured cell attached to the substrate via the laminin molecule or the fragment thereof, wherein the monolayer of the renal tubule epithelial cell or the renal tubule epithelial-like cell is maintained for 15 days or more,
wherein the laminin molecule or the fragment thereof is selected from any one of
Laminin-111 or a fragment thereof that adheres to the substrate at an amount of 0.15 μg/cm 2 or more,
Laminin-211 or a fragment thereof that adheres to the substrate at an amount of 0.52 μg/cm 2 or more,
Laminin-221 or a fragment thereof that adheres to the substrate at an amount of 0.34 μg/cm 2 ,
Laminin-311 or a fragment thereof that adheres to the substrate at a maximum amount after air drying of 0.5 μg/cm 2 or more,
Laminin-332 or a fragment thereof that adheres to the substrate at a maximum amount after air drying of 1.2 μg/cm 2 or more,
Laminin-421 or a fragment thereof that adheres to the substrate at an amount of 0.50 μg/cm 2 or more,
Laminin-511 or a fragment thereof that adheres to the substrate at an amount of 0.32 μg/cm 2 or more,
a variant of E8 region of Laminin-511 that adheres to the substrate at an amount of 0.15 μg/cm 2 or more, and
Laminin-521 or a fragment thereof that adheres to the substrate at an amount of 0.44 μg/cm 2 or more.

2. The cell support composite according to claim 1 , wherein the laminin 311 or the fragment thereof adheres to the substrate at an amount of 0.15 μg/cm 2 or more.

3. The cell support composite according to claim 1 ,
wherein the cultured cell has formed a confluent single layer without being substantially multilayered.

4. A method for producing the cell support composite of claim 1 , comprising:
coating at least a part of a substrate formed of an artificial material with a laminin molecule or a fragment thereof;
seeding a renal tubule epithelial cell or a renal tubule epithelial-like cell which is a cultured cell on the laminin molecule or the fragment thereof that has adhered to the substrate; and
culturing the cultured cell to give a single layer structure of the cultured cell,
wherein the coating with the laminin molecule or the fragment thereof comprises any one of:
coating with Laminin-111 at a concentration of 3.0 mg/ml or more to give an adhesion amount of 0.15 mg/cm 2 or more;
coating with Laminin-211 at a concentration of 10.0 mg/ml or more to give an adhesion amount of 0.52 mg/cm 2 or more;
coating with Laminin-221 at a concentration of 10.0 mg/ml or more to give an adhesion amount of 0.34 mg/cm 2 or more;
coating with Laminin-311 at a concentration of 3.0 mg/ml or more to give a maximum adhesion amount after air drying of 0.5 mg/cm 2 or more;
coating with Laminin-332 at a concentration of 5.0 mg/ml or more to give a maximum adhesion amount after air drying of 1.2 mg/cm 2 or more;
coating with Laminin-421 at a concentration of 5.0 mg/ml or more to give an adhesion amount of 0.50 mg/cm 2 or more;
coating with Laminin-511 at a concentration of 10.0 mg/ml or more to give an adhesion amount of 0.32 mg/cm 2 or more;
coating with a variant of E8 region of Laminin-511 at a concentration of 1.4 mg/ml or more to give an adhesion amount of 0.15 mg/cm 2 or more; and
coating with Laminin-521 at a concentration of 5.0 mg/ml or more to give an adhesion amount of 0.44 mg/cm 2 or more.

5. The method for producing a cell support composite according to claim 4 , wherein the coating with the laminin molecule or the fragment thereof comprises any one of:
coating with Laminin-311 at a concentration of 4.0 mg/ml or more to give an adhesion amount of 0.15 mg/cm 2 or more;
coating with Laminin-511 at a concentration of 10.0 mg/ml or more to give an adhesion amount of 0.32 mg/cm 2 or more;
coating with a variant of E8 region of Laminin-511 at a concentration of 1.4 mg/ml or more to give an adhesion amount of 0.15 mg/cm 2 or more; and
coating with Laminin-521 at a concentration of 5.0 mg/ml or more to give an adhesion amount of 0.44 mg/cm 2 or more.

6. The method for producing a cell support composite according to claim 4 , wherein the cultured cell has formed a confluent single layer without being substantially multilayered.

Assignees

• Nikkiso Co Ltd

Inventors

• Kitagawa Fumihiko

Classifications

• C12M25/06 Primary

  Plates; Walls; Drawers; Multilayer plates · CPC title

• C12N5/0068

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12N2533/52

  Fibronectin; Laminin · CPC title

• C12M25/00

  Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings · CPC title

• C07K14/705

  Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)} · CPC title